Quickchange site directed mutagenesis
Posted 05 May 2004 - 09:55 PM
Any tips? I'm not using the kit... I plan to purchase the Pfu Ultra and DpnI seperately.
Hints/suggestions greatly appreciated!
Posted 05 May 2004 - 11:06 PM
The strategy we've used for designing primers for situations for yours is just using the web utility to design a single-mutation primer, and then adding the second mutation to that and elongating the primer on the appropriate side.
Edited by btavshan, 05 May 2004 - 11:15 PM.
Posted 06 May 2004 - 07:19 AM
I had one more question... How big of a plasmid were you able to mutate? My plasmid is around 13kb and if I recall, stratgene's quickchange kit is for plasmids <8kb.
Also, did you ever encounter errors (i.e. pfu induced mutations?).